RNA-seq for V. amurensis
First-strand cDNA was synthesised using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H). Second strand cDNA synthesis was performed using DNA Polymerase I and RNase H. Remaining overhangs were converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3 ends of DNA fragments, adaptors with hairpin loop structure were ligated to prepare for hybridisation. To select cDNA fragments of preferentially 370 ~ 420 bp in length, the library fragments were purified with Agencourt AMPure XP (A63880/1/2, Beckman Coulter, Massachusetts, USA). PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers, and Index (X) Primer. Lastly, PCR products were purified with the AMPure XP system, and library quality was assessed on the Agilent Bioanalyzer 2100 system (2100, Agilent, USA). The index-coded samples were clustered on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (PE-401-3001, Illumia, USA) according to the manufacturers instructions. After cluster generation, the library preparations were sequenced on an Illumina Novaseq platform, and 150 bp paired-end reads were generated.